Primary cilia are nonmotile microtubule structures that assemble from basal bodies by a process called intraflagellar transport (IFT) and are associated with several human diseases. Here, we show that the centrosome protein pericentrin (Pcnt) colocalizes with IFT proteins to the base of primary and motile cilia. Immunogold electron microscopy demonstrates that Pcnt is on or near basal bodies at the base of cilia. Pcnt depletion by RNA interference disrupts basal body localization of IFT proteins and the cation channel polycystin-2 (PC2), and inhibits primary cilia assembly in human epithelial cells. Conversely, silencing of IFT20 mislocalizes Pcnt from basal bodies and inhibits primary cilia assembly. Pcnt is found in spermatocyte IFT fractions, and IFT proteins are found in isolated centrosome fractions. Pcnt antibodies coimmunoprecipitate IFT proteins and PC2 from several cell lines and tissues. We conclude that Pcnt, IFTs, and PC2 form a complex in vertebrate cells that is required for assembly of primary cilia and possibly motile cilia and flagella.
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30 August 2004
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August 30 2004
Pericentrin forms a complex with intraflagellar transport proteins and polycystin-2 and is required for primary cilia assembly
Agata Jurczyk,
Agata Jurczyk
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Adam Gromley,
Adam Gromley
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Sambra Redick,
Sambra Redick
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Jovenal San Agustin,
Jovenal San Agustin
2Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01605
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George Witman,
George Witman
2Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01605
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Gregory J. Pazour,
Gregory J. Pazour
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Dorien J.M. Peters,
Dorien J.M. Peters
3Leiden University Medical Center, Department of Human Genetics, 2333 AL Leiden, Netherlands
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Stephen Doxsey
Stephen Doxsey
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Agata Jurczyk
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
Adam Gromley
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
Sambra Redick
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
Jovenal San Agustin
2Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01605
George Witman
2Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01605
Gregory J. Pazour
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
Dorien J.M. Peters
3Leiden University Medical Center, Department of Human Genetics, 2333 AL Leiden, Netherlands
Stephen Doxsey
1Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
Address correspondence to Stephen Doxsey, Dept. of Molecular Medicine, University of Massachusetts Medical School, 373 Plantation Street, Biotech II, Suite 210, Worcester, MA 01605. Tel.: (508) 856-1613. Fax: (508) 856-4289. email: [email protected]
Abbreviations used in this paper: DIC, differential interference contrast; IFT, intraflagellar transport; PC2, polycystin-2; Pcnt, pericentrin; RPE1, retinal pigmented epithelial cell line 1; siRNA, small interfering RNA.
Received:
May 04 2004
Accepted:
July 13 2004
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2004
J Cell Biol (2004) 166 (5): 637–643.
Article history
Received:
May 04 2004
Accepted:
July 13 2004
Citation
Agata Jurczyk, Adam Gromley, Sambra Redick, Jovenal San Agustin, George Witman, Gregory J. Pazour, Dorien J.M. Peters, Stephen Doxsey; Pericentrin forms a complex with intraflagellar transport proteins and polycystin-2 and is required for primary cilia assembly . J Cell Biol 30 August 2004; 166 (5): 637–643. doi: https://doi.org/10.1083/jcb.200405023
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